JP6849390B2 - Joint stiffness improving agent - Google Patents

Description

The present invention relates to a joint stiffness improving agent.

Joint stiffness refers to a condition in which a joint is stiff and difficult to move at the beginning of movement. Joint stiffness is mainly caused by inflammation of the joints and tendons around the joints and appears in the joints of the whole body, but it is often felt in the hands and feet after not moving the joints for a long time, especially when waking up in the morning.
Even if you feel stiffness, you may be able to move the entire range of motion of the joint, and since the stiffness of the joint gradually diminishes, the symptoms are usually alleviated by moderately moving or warming the joint. It is planned. In addition, joint stiffness caused by autoimmune diseases such as rheumatoid arthritis and systemic lupus erythematosus is improved by treating the disease with drug therapy or the like.

On the other hand, milk-fat Globule Membrane (MFGM) is a membrane component that covers milk fat globules secreted from the mammary gland, and is abundant in milk complex lipid-rich fractions such as buttermilk and butterserum. It is known to be included (Non-Patent Document 1). It has been reported that the milk fat globules not only have a function of dispersing fat in milk, but also have a physiological function such as a motor function improving action and a muscular strength improving action in mice (Patent Document 1). In addition, it has been reported that the milk fat globules have an improving effect on stiff shoulders and tripping during walking (Patent Documents 2 and 3).
However, there are no reports on the effect of milk fat globules on joint stiffness.

Japanese Unexamined Patent Publication No. 2010-59155 Japanese Unexamined Patent Publication No. 2016-138508 Japanese Unexamined Patent Publication No. 2016-13859

Shin Miura, FOOD STYLE21, 2009

The present invention relates to providing a pharmaceutical product, a quasi-drug or a food product useful for improving joint stiffness, or a material or preparation that can be blended therein.

The present inventor has made a diligent study in view of the above problems, and found that ingestion of the milk fat globules reduces joint stiffness, and the milk fat globules are useful for improving joint stiffness.

That is, the present invention provides a joint stiffness improving agent containing a milk fat globules membrane as an active ingredient.
The present invention also provides a food for improving joint stiffness containing a milk fat globules as an active ingredient.

According to the present invention, the stiffness of joints such as knee joints can be improved.

A graph showing the effect of improving the stiffness of the knee joint.

The milk fat globules membrane used in the present invention is defined as a membrane covering the milk fat globules and a mixture of the components constituting the membrane. The milk fat globules have abundant eating experience and are highly safe.
It is known that the milk fat globules are generally composed of about half of the dry weight of lipids, and the lipids include triglyceride, phospholipids, and sphingolipids (Shin Miura, FOOD STYLE21, 2009 and Keenan TW, Applied Science Publicers, 1983, pp89-pp130). It is known that phospholipids include sphingolipids such as sphingomyelin (SM) and glycerophospholipids such as phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylserine (PS).
Further, it is known that a glycoprotein called milk mutin is contained as a component other than lipid (Mather, Biochim Biophys Acta, 1978).

From the viewpoint of physiological effects, the milk fat globules used in the present invention preferably have a lipid content of 10% by mass or more, further 20% by mass or more, and further 30% by mass or more. Further, from the viewpoint of flavor and handling, it is preferably 100% by mass or less, further 90% by mass or less, and further preferably 60% by mass or less.

From the viewpoint of physiological effects, the milk fat globules have a phospholipid content of 5% by mass or more, further 8% by mass or more, further 10% by mass or more, and further 15% by mass or more. It is preferable, and from the viewpoint of flavor and handling, it is preferably 100% by mass or less, further 85% by mass or less, further 70% by mass or less, and further 60% by mass or less.

From the viewpoint of physiological effects, the content of sphingomyelin (SM) in the milk fat globules is preferably 1% by mass or more, further 2% by mass or more, and further 3% by mass or more. Further, from the viewpoint of flavor and handling, it is preferably 50% by mass or less, further 30% by mass or less, further 25% by mass or less, and further 20% by mass or less.
From the same point of view, the content of sphingomyelin in the total phospholipids of the milk fat globules is preferably 3% by mass or more, further 5% by mass or more, further 10% by mass or more, and further 15% by mass or more. , 50% by mass or less, further 40% by mass or less, further 35% by mass or less, and further preferably 30% by mass or less.
In the present specification, the content of lipids and phospholipids in the milk fat globules, and the content of sphingomyelin in the total phospholipids of the milk fat globules are the mass ratio of the milk fat globules to the dried product. To do.

The milk fat globules can be obtained from raw milk by a known method such as a centrifugation method or an organic solvent extraction method. For example, the method for preparing a milk fat globules film described in JP-A-3-47192 can be used. Further, the methods described in Japanese Patent No. 3103218 and Japanese Patent Application Laid-Open No. 2007-89535 can be used.
Further, those whose purity has been increased by purification by a method such as dialysis, ammonium sulfate fractionation, gel filtration, isoelectric point precipitation, ion exchange chromatography, solvent fractionation and the like may be used.
The form of the milk fat globules is not particularly limited, and may be liquid, semi-solid (paste, etc.), solid (powder, solid, granules, etc.) at room temperature (15 to 25 ° C.), and these may be used alone. Although it may be used in combination of two or more types, it is preferably in a solid state (powder).

Examples of the raw material milk for the milk fat globules film include milk and goat milk. Of these, milk is preferable because it has abundant eating experience and is inexpensive. In addition, raw milk includes milk such as raw milk, whole milk powder and processed milk, as well as dairy products, and examples of dairy products include buttermilk, butter oil, butter salem, whey protein concentrate (WPC) and the like. Be done.
Buttermilk is obtained when producing butter grains from a cream obtained by centrifuging milk or the like, and since the buttermilk contains a large amount of milk fat globules, buttermilk is used as the milk fat globules. You may use it as it is. Similarly, since the butter salem produced during the production of butter oil also contains a large amount of milk fat globules, the butter salem may be used as it is as the milk fat globules.

As the milk fat globules film, a commercially available product can also be used. Examples of such commercially available products include "BSCP" from Megre Japan Co., Ltd., "Milk Ceramide MC-5" from Snow Brand Milk Products Co., Ltd., and "Phospholipid Concentrate Series (500, 700)" from New Zealand Milk Products Co., Ltd.

As shown in Examples below, ingestion of the milk fat globules film improved the stiffness of the knee joint that occurs when waking up, so that the milk fat globules membrane has an effect of significantly improving the stiffness of the joints.
Therefore, the milk fat globules can be a joint stiffness improving agent useful for improving joint stiffness, and can also be used to produce a joint stiffness improving agent. That is, the milk fat globules can be applied to humans who are concerned about joint stiffness and can be used to improve joint stiffness.
As used herein, the term "joint" refers to all connections between bones, of which movable connections are made. In the present invention, the joint is preferably a butterfly joint, a spiral joint, a saddle joint, an elliptical joint, a condylar joint, a ball joint, a flat joint, a half joint, a acetabular joint, and more preferably a knee joint or an interphalangeal joint. , Lumbar facet joints, shoulder joints, elbow joints, hip joints, and more preferably knee joints.
"Joint stiffness" is a symptom that the joint is stiff and difficult to move at the beginning of movement. This stiffness often appears when the joints are not moved for a long time due to sleep or maintaining the same posture, and the sensation fades and disappears shortly after it occurs. Stiffness also often manifests itself as a symptom before joint pain and swelling.
"Amelioration" means improvement of a symptom or condition, prevention, suppression or delay of exacerbation of the symptom or condition, or reversal, prevention, suppression or delay of progression of the symptom or condition.
"Use" can be administration or ingestion to a human and may be therapeutic or non-therapeutic use. "Non-therapeutic" is a concept that does not include medical practice, that is, a concept that does not include a method of operating, treating or diagnosing a human, more specifically, a doctor or a person who has been instructed to perform surgery on a human. , A concept that does not include a method of performing treatment or diagnosis.

The joint stiffness improving agent of the present invention is a drug, quasi-drug or food that exerts a joint stiffness improving effect when ingested or administered to animals including humans, and the joint stiffness improving agent is the said drug. , Can be a material or formulation used in combination with quasi-drugs or foods.

The foods include foods (foods for specified health use, foods with functional claims) that have the concept of improving joint stiffness and are permitted to be labeled to that effect as necessary. Since these foods are foods for which functional labeling is permitted, they can be distinguished from general foods.

Examples of the administration form of the above-mentioned pharmaceutical products (including quasi-drugs, the same applies hereinafter) include oral administration with tablets, capsules, granules, powders, troches, syrups and the like.
Formulations of these various dosage forms are the milk fat globules of the invention, or other pharmaceutically acceptable carriers, such as excipients, binders, bulking agents, disintegrants, surfactants, glides. It can be prepared by appropriately combining a swamp, a dispersant, a buffer, a preservative, a flavoring agent, a fragrance, a coating agent, a carrier, a diluent and the like, and medicinal ingredients other than the milk fat globules coating.
Among them, the preferred dosage form is a solid preparation for oral administration, and tablets are preferable.

The content of the milk fat globules in the pharmaceutical product (in terms of dry matter) is generally 0.01% by mass or more, preferably 0.1% by mass or more, more preferably 0.2% by mass or more, and further preferably. Is 1% by mass or more, preferably 90% by mass or less, more preferably 80% by mass or less, still more preferably 70% by mass or less, still more preferably 60% by mass or less.

The form of the food may be solid, semi-solid or liquid, for example, eating and drinking of soft drinks, tea-based beverages, coffee beverages, fruit juice beverages, carbonated beverages, jellies, wafers, biscuits, bread, noodles, sausages and the like. In addition to various food compositions such as products and nutritional foods, further, nutritional supplement compositions in the same form as the above-mentioned orally administered preparations (solid preparations such as tablets, capsules and troches) can be mentioned. Among them, solid preparations are preferable, and tablets are more preferable.

Foods of various forms include milk fat globules or other food materials, solvents, softeners, oils, emulsifiers, preservatives, acidulants, sweeteners, bitterness agents, fragrances, stabilizers, colorants, oxidation. It can be prepared by appropriately combining an inhibitor, a moisturizing agent, a thickener, an active ingredient other than the butterfat bulb film, and the like.

The content of the milk fat globules in the food (in terms of dry matter) varies depending on the form of use, but in the form of a beverage, it is preferably 0.001% by mass or more, more preferably 0.01% by mass or more, still more preferably Is 0.1% by mass or more, preferably 3% by mass or less, more preferably 2% by mass or less, still more preferably 1% by mass or less.

In the form of solid foods such as tablets and processed foods, the content of the milk fat globules (in terms of dry matter) is preferably 0.05% by mass or more, more preferably 0.1% by mass or more, still more preferably 0. It is 2% by mass or more, preferably 90% by mass or less, more preferably 80% by mass or less, still more preferably 70% by mass or less, still more preferably 60% by mass or less.

The dose or ingestion of the joint stiffness improving agent of the present invention may vary depending on the body weight, sex, age, condition or other factors of the subject to be administered or ingested. The dose, route, interval of administration, and the amount and interval of ingestion can be appropriately determined by those skilled in the art, but usually as a milk fat globules (dry matter equivalent) per day for one adult (60 kg). It is preferably 0.1 g or more, more preferably 0.3 g or more, still more preferably 1 g or more, and preferably 30 g or less, more preferably 20 g or less, still more preferably 10 g or less.
In addition, usually, as sphingomyelin per day for one adult (60 kg), the amount of sphingomyelin is preferably 10 mg or more, more preferably 20 mg or more, further preferably 40 mg or more, and preferably 1500 mg or less, more preferably. It is 1000 mg or less, more preferably 500 mg or less, still more preferably 250 mg or less.
In the present invention, it is preferable to administer or ingest such an amount once to a plurality of times a day.

The above formulations may be administered or ingested according to any plan.
The administration or ingestion period is not particularly limited, but it is preferable to administer or ingest repeatedly and continuously, more preferably to administer or ingest continuously for 5 days or more, and to administer or ingest continuously for 15 days or more. More preferred.

The subject to be administered or ingested is not particularly limited as long as it is a person who needs or desires to reduce joint stiffness and improve stiffness, but administration or ingestion in a person who is aware of joint stiffness is effective. is there.

[Preparation of tablets]
Tablets having the compositions shown in Table 1 below (Example 1: 353 mg / tablet, Comparative Example 1: 450 mg / tablet) were prepared according to the Japanese Pharmacopoeia (general formulation "tablets").

The milk fat globules membrane (MFGM) used was prepared from milk.
The water content of MFGM was 3.6% by mass. The composition of MFGM was 11.3% by mass of carbohydrates, 25.1% by mass of lipids, and 53.6% by mass of proteins in terms of dry matter. In addition, the content of phospholipid in MFGM was 16.6% by mass in terms of dry matter, and the content of sphingomyelin was 3.6% by mass.

The above analysis of MFGM was performed as follows.
(1) Protein analysis The amount of protein was determined using the Kjeldahl method with a nitrogen-protein conversion coefficient of 6.38.

(2) Analysis of lipid The amount of lipid was determined by the acid decomposition method. 1 g of the sample was weighed, hydrochloric acid was added to decompose the sample, diethyl ether and petroleum ether were added, and the mixture was stirred and mixed. The ether mixed liquid layer was taken out and washed with water. After distilling off the solvent and drying, the amount of lipid was determined by weighing.

(3) Analysis of carbohydrates The amount of carbohydrates was determined by removing the amount of protein, the mass of lipids, the amount of ash, and the amount of water in the sample from the mass of the sample. The amount of ash was determined by the direct ashing method (the sample was ashed at 550 ° C. and weighed), and the amount of water was determined by the atmospheric heating drying method (drying at 105 ° C. for 4 hours and measuring the weight).

(4) Analysis of phospholipids 1 g of a sample is weighed and homogenized in 150 mL, 100 mL, and 20 mL of a 2: 1 (V / V) mixed solution of chloroform and methanol, and then 0.88 mass% (W / V) aqueous potassium chloride solution. 93 mL was added and left overnight at room temperature. After dehydration filtration and solvent distillation, chloroform was added to bring the total volume to 50 mL. Of that, 2 mL was separated, and after distilling off the solvent, it was incinerated by heat treatment at 550 ° C. for 16 hours. After dissolving the ash in 5 mL of a 6 M hydrochloric acid aqueous solution, distilled water was added to bring the total volume to 50 mL. 3 mL was separated, 5 mL of molybdenum blue coloring reagent, 1 mL of 5% by mass (W / V) ascorbic acid aqueous solution and distilled water were added to make the total volume 50 mL, and the absorbance at 710 nm was measured. The amount of phosphorus was determined from a calibration curve using potassium dihydrogen phosphate, and the value obtained by multiplying the amount of phosphorus by 25.4 was defined as the amount of phospholipid.

(5) Analysis of sphingomyelin Weigh 1 g of a sample, homogenize it in 150 mL, 100 mL, and 20 mL of a 2: 1 (V / V) mixture of chloroform and methanol, and then 0.88% by mass (W / V) aqueous potassium chloride solution. 93 mL was added and left overnight at room temperature. After dehydration filtration and solvent distillation, chloroform was added to bring the total volume to 50 mL. Of that, 10 mL was dispensed and added to the silica cartridge column. The column was washed with 20 mL of chloroform, phospholipids were eluted with 30 mL of methanol, the solvent was distilled off, and the column was dissolved in 1.88 mL of chloroform. 20 μL is loaded on the silica gel thin layer plate, and tetrahydrofuran: acetone: methanol: water = 50: 20: 40: 8 (V / V) as the one-dimensional developing solvent, and chloroform: acetone: methanol: acetic acid: water as the two-dimensional developing solvent. Two-dimensional development was performed using = 50: 20: 10: 15: 5 (V / V). A ditomer reagent was sprayed on the developed thin-layer plate, spots of sphingomyelin were scraped off, 2 mL of a 3% by mass (V / V) nitric acid-containing perchloric acid solution was added, and then heat treatment was performed at 170 ° C. for 3 hours. After adding 5 mL of distilled water, 5 mL of the molybdenum blue coloring reagent and 1 mL of a 5 mass% (W / V) ascorbic acid aqueous solution and distilled water were added to make the total volume 50 mL, and the absorbance at 710 nm was measured. The amount of phosphorus was determined from a calibration curve using potassium dihydrogen phosphate, and the value obtained by multiplying the amount of phosphorus by 25.4 was defined as the amount of sphingomyelin.

[Double-blind, randomized, parallel design comparative verification test]
1. 1. Subjects and test method 16 healthy men and women in their 60s who had a feeling of stiffness in the knee joint when waking up in the morning were divided into two groups, the MFGM group (7 people) and the placebo group (9 people), and were divided into the MFGM group. Ingested the tablets of Example 1 containing MFGM (10 tablets in total) and the tablets of Comparative Example 1 containing no MFGM (7 tablets in total) in the placebo group every day for 6 weeks at a desired timing.

2. Evaluation of knee joint stiffness improvement effect Evaluation of knee joint stiffness improvement effect is based on the "stiffness" of the knee joint before the start of the test (week 0), 6 weeks after the start of the test, after waking up in the morning, and when starting to move. The condition was evaluated by interviewing. The evaluation criteria are "1: not stiff", "2: slightly stiff", "3: moderately stiff", "4: fairly stiff", and "5: severely stiff", each of which is in a state of several days. The score value that seems to be the most applicable was decided.

3. 3. Results All the subjects were selected as the final analysis subjects, and the difference (Δ value) between each group and the initial value was calculated for the score value after 6 weeks and shown in FIG.
The obtained values are shown as mean ± standard deviation.
As a result, it was confirmed that the stiffness of the knee joint before the start of the test and after the end of the test was reduced in the MFGM group as compared with the placebo group, and the stiffness of the joint was improved by ingestion of the milk fat bulb membrane.

Claims (11)

A joint stiffness improving agent containing milk fat globules as an active ingredient (excluding use as a joint stiffness improving agent due to rheumatoid arthritis) . The joint stiffness improving agent according to claim 1, wherein the milk fat globules film contains 5 to 100% by mass of phospholipids. The joint stiffness improving agent according to claim 1, wherein the milk fat globules film contains 1 to 50% by mass of sphingomyelin. The joint stiffness improving agent according to any one of claims 1 to 3, wherein 10 to 1500 mg of sphingomyelin is administered or ingested daily per adult. The joint stiffness improving agent according to any one of claims 1 to 4, wherein the joint is a knee joint. The joint stiffness improving agent according to any one of claims 1 to 5, wherein the dosage form is a solid preparation for oral administration. Foods for improving joint stiffness containing milk fat globules as an active ingredient (excluding use as foods for improving joint stiffness due to rheumatoid arthritis) . The food for improving joint stiffness according to claim 7, wherein the milk fat globules film contains 5 to 100% by mass of phospholipids. The food for improving joint stiffness according to claim 7, wherein the milk fat globules film contains 1 to 50% by mass of sphingomyelin. The food for improving joint stiffness according to any one of claims 7 to 9, wherein the joint is a knee joint. The food for improving joint stiffness according to any one of claims 7 to 10, wherein the dosage form is a solid preparation for oral administration.

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